Germ Tube Test Procedure MoH/DGQAC/P&P/002/Vers.

Effective Date: Month/ Year

Review Date: Month/ Year

Institution Name: Ibri regional Hospital

Document Title: Germ Tube Test Procedure

Approval Process

Name Title Institution Date Signature

Written by Asma Al- Lab Ibri

Yaqoobi Technicin Regional
Hospital

Reviewed by

Validated by

Approved by
 Germ Tube Test Procedure MoH/DGQAC/P&P /002/Vers.
Effective Date: Month/ Year
Review Date: Month/ Year

Contents Table:
Acronyms.................................................................................................................1
1. Introduction.....................................................................................................2
2. Scope...............................................................................................................2
3. Purpose............................................................................................................2
4. Policy..............................................................................................................2
5. Procedure........................................................................................................2
7. Limitations......................................................................................................4
9. Document History and Version Control.........................................................5
10. References.......................................................................................................5
Acronyms:

°C Degree Celsius

C. Candida

GT Germ Tube

KOH Potassium Hydroxide

ml Milliliter

MOH Ministry of Health

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 Germ Tube Test Procedure MoH/DGQAC/P&P/002/Vers.
Effective Date: Month/ Year
Review Date: Month/ Year

1. Introduction
The germ- tube test is the most generally accepted and economical method
used in the clinical laboratory for the identification of yeasts. This test
usually provides a sufficient identification within 3 hours.

Germ tube (GT) formation was first reported by Reynolds and Braude in
1956. When Candida is grown in human or sheep serum at 37°C for 3 hours,
they forms a germ tubes, which can be detected with a wet KOH films as
filamentous outgrowth extending from yeast cells. It is positive for Candida
albicans and Candida dubliniensis. Approximately 95 – 97% of Candida
albicans isolated develop germ tubes when incubated in a proteinaceous
media.

2. Scope
This document is applicable to the Department of Laboratory Services, Ibri
Regional Hospital, MOH.

3. Purpose
The purpose of the document is to describe the policy and procedure of Germ
tube.

4. Policy
All rotating technical staffs posted in the Microbiology Department are
permitted to perform this test with guidance from Supervisory staff as and
when necessary.

5. Procedure
The main use of the test is to differentiate Candida albicans from other yeast.

5.1. Principle
Formation of germ tube is associated with increased synthesis of protein and
ribonucleic acid. Germ Tube solutions contains tryptic soy broth and fetal

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bovine serum, essential nutrients for protein synthesis. It is lyophilized for
stability. Germ tube is one of the virulence factors of Candida albicans. This is
a rapid test for the presumptive identification of C. albicans.

5.2. Material and Reagents Required

5.2.1. Sheep or human serum
5.2.2. Yeast to be examined
5.2.3. Small test tubes
5.2.4. Pasteur pipette
5.2.5. Plain slide and cover slip

5.3. Testing Procedure

5.3.1. Put 0.5 ml of sheep or human serum into a small tube.

Note: Fetal bovine serum can also be used instead of human serum.
5.3.2. Using a Pasteur pipette, touch a colony of yeast and gently emulsify it in
the serum.
Note: Too large of an inoculum will inhibit germ tube formation.
5.3.3. Incubated the tube at 37°C for 2 to 4 hours.
5.3.4. Transfer a drop of the serum to a slide for examination.
5.3.5. Cover slip and examine microscopically under low and high
power objectives.

5.4. Results

5.4.1. Positive Test: A short hyphal (filamentous) extension arising laterally

from a yeast cell. Germ tube is half the width and 3 to 4 times the
length of the yeast cell and there is no presence of nucleus.

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 5.4.2. Negative Test: No hyphal (filamentous) extension arising from a
yeast cell or a short hyphal extension constricted at the point of origin.

6. Quality Control

6.1. Positive Control: C. albicans

6.2. Negative Control: C. tropicalis and C. glabrata.

7. Limitations
7.1. C. tropicalis may form early pseudohyphae which may be
falsely interpreted as germ tubes.
7.2. The yeast formerly named Candida stellatoidea also produces germ tubes;
however, it has been combined with C. albicans and no longer exists as
separate species.
7.3. This test is only part of the overall scheme for identification of
yeasts. Further testing is required for definite identification.

8. Responsibilities
Performing the germ tube test and follow up is responsibility of Microbiology
section staff.

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9. Document History and Version Control

Document History and Version Control

Version Description of Amendment Author Review

Date

01 Initial Release

02

03

04

05

Written by Reviewed by Approved by

10. References:

Title of book/ journal/ articles/ Website Author Year of Page

publication

https://microbiologyinfo.com/germ-tube-test- 2014
principle-procedure-results-interpretation-and-
limitations/

Bailey and Scott’s DIAGNOSTIC MICROBILIOGY Bailey and 700-

( 12 Edition)
th Scott’s 701

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